AFFINITY-CHROMATOGRAPHY OF SULFATED POLYSACCHARIDES SEPARATELY FRACTIONATED ON ANTITHROMBIN-III AND HEPARIN COFACTOR-II IMMOBILIZED ON CONCANAVALIN-A-SEPHAROSE

Three sulphated polysaccharides, dermatan sulphate, fucan and heparin, were fractionated according to their affinity towards antithrombin II I (ATIII) and heparin cofactor II (HCII), the two main physiological t hrombin (IIa) inhibitors. Both inhibitors were immobilized on concanav alin A-Sepharose, which binds to the glycosylated chains of the protei ns while the protein-binding site for the polysaccharide remains free. Each polysaccharide was fractionated into bound and unbound fractions either for ATIII or HCII. The eluted fractions were tested for their ability to catalyse ATIII/Ila and HCII/IIa interactions. The possible presence of a unique binding site for ATIII and HCII, on each sulphate d polysaccharide, was also studied.